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Biomedical subjects

J M Wilkinson

Publications and source records attributed to J M Wilkinson.

At least 91 records · Page 5Linked to original sources
PubMed1993

Immunohistochemical identification of leucocyte populations in normal tissue and inflamed synovium of the rabbit.

A panel of monoclonal antibodies which recognizes well-defined markers of rabbit leucocytes has been used in an immunohistochemical survey of normal rabbit spleen, thymus, and liver, and of inflamed synovial tissue. The results demonstrate that these antibodies are well suited for the identification of leucocytes in frozen sections of rabbit tissues and that T lymphocytes, macrophages, and neutrophils can be readily distinguished. Antibodies to CD44 differentiate between medullary and cortical thymocytes, with the cortical cells being almost entirely negative for this marker. Antibodies to RLA class II show that periportal lymphocytes in the liver express RLA-DR but not RLA-DQ. The type A cells of the synovial lining from inflamed joint tissue are stained by antibodies that recognize macrophages in a manner similar to that found in the human synovial lining.

Animals↗
PubMed1993

Monoclonal antibodies to cytoskeletal proteins: an immunohistochemical investigation of human colon cancer.

Monoclonal antibodies raised to a number of microfilament-associated proteins were shown to recognize the appropriate proteins in extracts from human colon tissue. They were then used in an immunohistochemical study of normal colonic mucosa, adenomas, and adenocarcinomas. A strong reaction was seen in stromal cells within the tumours (both adenomas and adenocarcinomas) when frozen sections were stained with antibodies to filamin and caldesmon. In addition, a similar reaction was seen in the adenocarcinomas when stained with antibodies to talin and gelsolin. We believe that immunohistochemical staining with these antibodies reveals a tumour-induced process in the surrounding cells, possibly related to a host response to tumours.

Adenocarcinoma↗
PubMed1993

The expression of CD11/CD18 molecules on rabbit leucocytes: identification of monoclonal antibodies to CD18 and their effect on cellular adhesion processes.

The monoclonal antibodies, L13/64 and RCN1/21, raised against rabbit leucocytes, have been shown, by sequential immunoprecipitation and immunoblotting, to react with the rabbit CD18 molecule. They recognise not only surface-expressed CD18 but also an intracellular form which appears to be partially glycosylated. The expression of the CD11 and CD18 glycoproteins on a wide variety of rabbit leucocyte populations has been investigated by flow cytometry, using these two monoclonal antibodies (Mabs), together with others which recognise human CD11 and CD18 proteins but cross-react with rabbit tissues. The distribution of these leucocyte integrin molecules has been shown to be similar to that observed in humans and determination of the N-terminal sequence of rabbit CD11b shows strong homology with human and mouse sequences. Of four anti-rabbit CD18 Mabs tested, only one, L13/64, has been shown to be capable of inhibiting the adhesion of fMLP-stimulated neutrophils to gelatin coated plastic and the homotypic aggregation of PMA-stimulated T cells, both of which assays have been shown to be CD18-dependent. RCN1/21 causes aggregation of unstimulated neutrophils, but it is not known whether this is due to cellular activation or agglutination.

Amino Acid Sequence↗
PubMed1993

Characterization of monoclonal antibodies against rabbit CD44: evidence of a role for CD44 in modulating synoviocyte metabolism.

The CD44 adhesion receptor is a hyaluronan-binding cell surface glycoprotein whose major isoform is a 95 kDa molecule with a wide tissue distribution. We have characterized two MAbs, W4/86 and RPN3/24, by immunochemistry, flow cytometry and fluorescence staining and demonstrated that they recognize the rabbit CD44 molecule. The results show that CD44 in the rabbit has a similar molecular size and cell distribution to human CD44. These antibodies have demonstrated for the first time, the presence of CD44 on synoviocytes. Preincubation of the rabbit synovial cell line HIG-82 with W4/86 inhibited its synthesis of PGE2 in response to autocrine synovial cytokines. These results indicate that the CD44 molecule may help to modulate synoviocyte metabolism.

Animals↗
PubMed1993

High technology to simplify the planning and delivery of radiotherapy.

Two techniques for the automatic selection of individual leaf positions of a Philips multi-leaf collimator are described. Target volumes are identified either on simulator images or on cross-sectional images from CT or MR scanners. The setting of each leaf is computed to position the beam edge to cover the target with an appropriate, user defined, margin. An important consideration in the development of the system was its robustness and so the applications initially implemented have been relatively simple, comprising single field, parallel opposed fields and coplanar 4 field box techniques. Attention has been paid to the overall integrity of the planning and treatment delivery process. Before treatment commences, the beam shapes, which have been generated by the computer and transferred to the MLC control computer over a local area network, are checked against a printed template representing each beam. All data used for planning is archived and is accessible for review or, if necessary, for treatment modification.

Humans↗
PubMed1993

Caffeine does not increase synthesis of heat shock proteins in rat embryos.

Caffeine exposure in utero in rats is known to result in intrauterine growth retardation and lowered birth weight as well as changes to behaviour and brain biochemistry. We have investigated whether caffeine's embryotoxicity is a result of the events associated with increased hsp synthesis, i.e., disruption to normal protein synthesis. Caffeine (30 mg/kg) was administered orally to pregnant rats as single or repeated doses. Embryos were removed 3 h after dosing on gestation day (GD) 9, 10, 11 and 12 and total embryonic protein and RNA analysed. There was no change in the mRNA or protein levels of hsp 88, 71/73, and 25 after acute or chronic treatment. To separate the direct effect of caffeine from those mediated through the mother, whole rat embryo culture was used. Caffeine (50 micrograms/ml) for 90 min did not increase hsp 88, 73 or 25 mRNA levels in 9.5, 10.5 and 11.5 GD cultured embryos. We conclude that in vivo or in vitro treatment of 9-12 GD rat embryos with moderate to high doses of caffeine does not increase the synthesis of the major mammalian hsps. Hence, hsp induction is unlikely to play a role in the embryotoxic actions of caffeine.

Animals↗
PubMed1993

Immunohistochemical localisation of the 90, 70 and 25 kDa heat shock proteins in control and caffeine treated rat embryos.

Human and animal experimental data demonstrate that in utero exposure to caffeine results in intrauterine growth retardation and long-term behavioural and reproductive effects. We have suggested that the disruption of normal transcription and translation associated with the initiation of the heat shock response may be a possible mechanism of action of caffeine. This hypothesis was investigated using immunohistochemistry to determine whether an acute (3 h) dose of 30 mg/kg caffeine alters the distribution of hsp 90, 70 and 25 in 10.5-12.5 g.d. rat embryos. In the control embryos hsps 90 and 70 were distributed throughout the embryo with no areas of specific accumulation. Hsp 25 was localised to the developing myocardium of 10.5, 11.5 and 12.5 g.d. embryos and the myotome of 11.5 and 12.5 g.d. embryos. The appearance of hsp 25 was correlated with the onset of muscle fibre differentiation and it is suggested that hsp 25 is associated with cytoskeletal proteins. Following dosing with caffeine no change in the distribution of staining for hsp 90, 70 and 25 was found. These results strongly suggest that caffeine's mechanism of action does not involve initiation of the heat shock response.

Animals↗
PubMed1993

The efficacy of lead shielding in megavoltage radiotherapy.

The dose received by anatomical structures which are apparently shielded by lead blocks during a course of megavoltage radiotherapy may not be negligible. The dose has three main components; there is that due to radiation transmitted through the shielding block, that due to radiation scattered within the patient and that due to radiation scattered prior to reaching the patient and so circumventing the shield. A calculation method which separates the three components has been investigated and the results have been compared with measurements in a wide range of test situations for both cobalt-60 and 6 MV photon beams. The method of calculation predicts the dose behind the shielding block with acceptable accuracy in all the conditions investigated.

Cobalt Radioisotopes↗
PubMed1993

Localisation of pp60c-src to the surface membrane of human platelets.

In platelets, pp60c-src has been variously localised to cytosol, dense bodies, or plasma membrane and surface-connected canalicular membranes. We have investigated the localisation of pp60c-src in highly purified human platelet subcellular fractions isolated using density gradient centrifugation and continuous flow electrophoresis followed by quantitative immunoblotting. We have obtained fractions enriched in surface membranes and alpha granule membranes as assessed by the presence of the marker proteins gpIb, gpIIb, thrombospondin, fibrinogen and fibronectin. Quantitative immunoblotting demonstrated that most of the cellular pp60c-src co-purified with markers for surface membrane components and that unlike the situation in fibroblasts, platelet pp60c-src did not associate with non-granule intracellular membranes such as Golgi, endoplasmic reticulum and endosomes. A small subpopulation of the total cellular pp60c-src co-purified with granule components or dense body markers, however this occurred in proportion to the presence of surface membrane markers. Our results demonstrate that the majority of platelet pp60c-src is located on the cytoplasmic face of the plasma membrane in human platelets.

Blood Platelets↗
PubMed1993

Situational supports and job stress: a correlational study.

This correlational study provides information for creating a nursing work environment that supports coping with stress. Questionnaire-generated data were collected from registered staff nurses (N = 538) in two urban, acute care hospitals. Results of multiple regression analysis indicated that situational support variables (autonomy, control over practice, group cohesion, substantive exchange, manager consideration) correlated inversely with job stress. Predictor variables and demographics explained 37% of the variance in situational job stress [F(13, 442) = 19.74, P < .001] and 28% of the variance in personal job stress [F(13, 442) = 13.03, P < .001]. The variables may serve as situational supports for nurses coping with job stress.

Adaptation, Psychological↗
PubMed1992

Binding of human platelet glycoprotein Ib and actin to fragments of actin-binding protein.

Actin-binding protein (ABP) is degraded into fragments of 190 and 90 kDa by calpain. A monoclonal antibody (MAb TI10) against the 90 kDa fragment of ABP coprecipitated with the glycoprotein Ib (GP Ib) peak observed on crossed immunoelectrophoresis of Triton X-100 extracts of platelets prepared without calpain inhibitors. MAb PM6/317 against the 190 kDa fragment was not coprecipitated with the GP Ib peak under such conditions. The 90 kDa fragment was adsorbed on protein A agarose from extracts that had been preincubated with antibodies to GP Ib. This supports the idea that the GP Ib-ABP interaction resides in the 90 kDa region of ABP. GP Ib was sedimented with the Triton-insoluble actin filaments in trace amounts only, and only after high speed centrifugation (100,000 x g, 3 h). Both the 190 kDa and the 90 kDa fragments of ABP were sedimented with the Triton-insoluble actin filaments.

Actins↗
PubMed1992

Position of the fovea centralis with respect to the optic nerve head.

Examination of the ocular fundus via indirect ophthalmoscopy gives the clinician an impression of foveal position relative to the nerve head. In some patients, the fovea appears to be in an unusual position (i.e., ectopic): it may appear to be higher or lower than expected, or closer to or farther from the nerve head. There is little published quantitative information on this subject. The purpose of this study was to examine foveal position in a group of normal adult eyes, so that clinicians and other researchers will be able to determine on a more objective basis whether or not a given patient shows foveal ectopia. Using ocular fundus photographs for 446 normal adult eyes, we found the foveal center to be, on average, 6.11 degrees +/- 3.32 degrees below a horizontal line bisecting the nerve head. For a smaller sample of 66 eyes, we found the average distance between the nerve head and foveal centers to be 4.93 +/- 0.33 mm (right eye) and 4.88 +/- 0.36 mm (left eye). Correlations of these data for right and left eyes are also examined. Nerve head data for the group of 66 right eyes were also analyzed to yield dimensions of a best-fitting ellipse: the mean minor axis was 1.75 +/- 0.2 mm; the mean major axis 1.95 +/- 0.2 mm. Ectopia (heterotopia) of the fovea has been found in association with chorioretinitis, fibrous traction bands, and/or colobomas of the choroid and optic nerve (including anomalous insertion of the optic nerve), microcephalus, and microphthalmia. A number of separate cases with anomalous nerve heads and/or foveal positions are discussed in this paper.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1992

Computer-assisted generation of multi-leaf collimator settings for conformation therapy.

Techniques for the automatic set up of the individual leaf positions of a Philips multi-leaf collimator system to cover a defined target volume are described. Tumour outline data for multi-field treatments may be obtained from one of two techniques, either from simulator images or from cross-sectional computed tomography (CT) slices. In the first technique, simulator images are digitized directly from image intensifier video signals or from conventional film radiographs using a CCD camera. Corrections for image distortion are carried out before reformatting the digitized images to a common data structure. Target outlines are subsequently traced interactively on the digital image to create an outline file. In the second technique, target volumes are defined on several individual CT slices and these are then used to obtain projected graphical views from any desired angle. In both techniques, a scaled graphical representation of leaf positions is then displayed and set relative to the outer edge of the target outline. Both techniques allow interactive repositioning of single leaves when required, or the operator can specify a margin around the projected target volume. Leaf prescription data files are created and are transferred via a Decnet-OSI-Opennet network link to an Intel microcomputer, which is used to drive the device itself. Examples of both techniques are given.

Humans↗
PubMed1992

A cytotoxic rabbit T-cell line infected with a gamma-herpes virus which expresses CD8 and class II antigens.

A rabbit T-cell line, BJ-610, has been derived from a New Zealand White rabbit infected with Alcelaphine herpes virus-1, which has the characteristics of a lymphokine activated killer (LAK) cell. The surface phenotype of this cell line has been studied by flow cytometry, using a panel of monoclonal antibodies (mAb) to rabbit leucocyte surface markers, and compared with that of another rabbit T-cell line, RL-5, transformed with herpes virus ateles. The expression of a number of markers is common to the two lines; these include the rabbit analogues of CD11a/CD18, CD43, CD44 and CD45. Three antigens are expressed on BJ-610 but not RL-5. One of these is recognized by a mAb thought to recognize CD8, while a second is a class II R-DQ molecule. The third antigen is expressed on thymocytes, a subset of T cells, neutrophils and platelets but its molecular nature is unknown. These two cell lines should prove useful in preparing reagents which recognize subsets of rabbit T cells and for studying the mechanism of herpes virus-induced lymphoid cell deregulation.

Animals↗